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Diss Factsheets

Environmental fate & pathways

Biodegradation in water: screening tests

Administrative data

biodegradation in water: ready biodegradability
Type of information:
experimental study
Adequacy of study:
key study
Study period:
December 1994 - February 2005
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: study performed in accordance with the corresponding OECD-/METI-testing guidelines
Reason / purpose for cross-reference:
reference to same study

Data source

Reference Type:
study report
Report date:

Materials and methods

Test guideline
according to guideline
other: Japanese testing method
Principles of method if other than guideline:
The test was conducted according to the "method for Testing the Biodegradability of Chemical Substances by microorganisms" stipulated in the "Testing Methods for New Chemical Substances" (July 13, 1974, Kanpogyo No.5, Planning and Coordination Bureau, Environment Agency, Yakuhatu No.615, Pharmaceutical Affairs Bureau, Ministry of Health and Welfare, and 49 Kikyoku No.392, Basic Industries Bureau, Ministry of International Trade and Industry, Japan).
GLP compliance:

Test material

Constituent 1
Chemical structure
Reference substance name:
EC Number:
EC Name:
Cas Number:
Molecular formula:
Details on test material:
- Name of test material (as cited in study report): Valeronitrile
- Molecular formula: C5 H9 N
- Molecular weight: 83.13
- Physical state: liquid
- Expiration date of the lot/batch: not mentioned
- Stability under test conditions: stable
- Storage condition of test material: dark and cool place

Study design

Oxygen conditions:
Inoculum or test system:
other: mixture of ten sludges from Japanese sewage treatment plants and surface waters
Details on inoculum:
Sludge was sampled from ten different locations in Japan:
- Fukogawa city sewage plant (Sapporo-shi Hokkaido)
- Fukashiba industry sewage plant (Kashima-gun Ibaragi)
- Nakahama city sewage plant (Osaka-shi Osaka)
- Ochiai city sewage plant (Shinjuku-ku Tokyo)
- Kitakami river (Ishinomaki-shi Miyagi)
- Shinano river (Nishikanbara-gun Niigata)
- Yoshino river (Tokushima-shi Tokushima)
- Lake Biwa (Otsu-shi Shiga)
- Hiroshima bay (Hiroshima-shi Hiroshima)
- Dookai bay (Kitakyushu-shi Fukuoka)

Mixing of fresh and old activated sludge:
5 l of the filtrate of the supernatant of activated sludge in use at present was mixed with 500 ml each of the filtrate of the supernatant of newly collected sludges and mixture was cultured at pH 7.0 + 1.0 under the sufficient aeration (Prefiltered open air/was used for aeration).

About 30 minutes after ceasing the aeration of the sludge mixture, supernatant corresponding to about 1/3 of the whole volume was removed. Then an equal volume of dechlorination water was added to the remaining portion. This mixture was aerated, and then synthetic sewage (Glucose, peptone and monopotassium phosphate respectively were dissolved in dechlorinated water. Each concentration was 5 (W/V) % and the solution was adjusted to pH 7.0 + 1.0 with sodium hydroxide).

During the culturing, the appearance of the supernatant, setting of the sludge, formation of flock, pH, dissolved oxygen concentration in the solution and temperature were checked and necessary adjustments were made. Microflora in the activated sludge was microscopically observed and sludge with no abnormal symptoms was used for the test.
Duration of test (contact time):
28 d
Initial test substance concentration
Initial conc.:
100 other: ppm
Based on:
test mat.
Details on study design:
Standard test design as recommended by the test guideline.
Reference substance
Reference substance:
(Wako Pure Chemical Industries, Ltd., Lot No. KCG9315)

Results and discussion

Preliminary study:
% Degradation
% degradation (O2 consumption)
St. dev.:
Sampling time:
28 d
Details on results:
Percentage biodegradation
- by TOC: 99.2%
- by GC: 100.0%
- by BOD, when N was oxidized to NO2: 64.0%
- by BOD, when N was released as NH4: 81.3%

BOD5 / COD results

Results with reference substance:
Percentage biodegradation of Aniline by BOD after 28 days = 71.3%

Applicant's summary and conclusion

Validity criteria fulfilled:
Interpretation of results:
readily biodegradable
Based on a mean biodegradation rate of 64% (by BOD) after 28 days, the substance can be regarded as readily biodegradable.
Executive summary:

The test was performed in 1994/1995 in conformity with GLP and following the Japanese testing methods for screening of chemicals. A concentration of 100 ppm test item was tested over 28 days on a mixture of activated sludges from ten different sewage treatment plants and surface waters in Japan. The test item showed a biodegradation of: 64% by BOD based on NO2, 81.3% by BOD based on NH4, 99.2% by TOC and 100% by GC. The reference substance aniline showed a biodegradation of 71% after 28 days.